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Orly Weinreb, Martinus Adrianus Maria van Boekel, Ahuva Dovrat, Hans Bloemendal; Effect of UV-A Light on the Chaperone-like Properties of Young and Old Lens α-Crystallin. Invest. Ophthalmol. Vis. Sci. 2000;41(1):191-198.
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purpose. To study the damaging effect of UV-A irradiation on the chaperone-like
properties of α-crystallin and the subsequent recovery process of
young and old bovine lenses.
methods. Young and old bovine lenses were kept in organ culture. After 24 hours
of incubation they were irradiated with UV-A at 365 nm, and optical
quality measurements were performed during the experiments (192 hours).α
-Crystallin and αA1-, αA2-, αB1-, and αB2-crystallin
subunits were analyzed, separated by gel filtration and cation exchange
chromatography, respectively, after different culture times. Protein
patterns were obtained after two-dimensional (2-D) gel electrophoresis.
Chaperone-like activity was determined on the basis of insulin B-chain
and βL-crystallin aggregation assays. Aggregation of α-crystallin
was analyzed, tryptophan fluorescence measurements were performed, andα
-crystallin mRNA levels were determined.
results. The water-soluble α-crystallin obtained from old lenses compared with
young lenses after UV irradiation had decreased chaperone activity, a
higher molecular weight, and increased loss of tryptophan fluorescence.
Moreover, α-crystallin mRNA virtually disappeared, whereas extra
spots on the 2-D protein pattern appeared, possibly because of
conclusions. α-Crystallin obtained from old lenses is more affected by irradiation
than α-crystallin derived from young lenses. Moreover, it appeared
that αB-crystallin from UV-treated old lenses compared with control
lenses was less susceptible to UV-A than αA-crystallin. It may well
be that αB-crystallin protects αA-crystallin in
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